Apparatus and method for implementing particle modification

Abstract

The application provides a device and a method for realizing particle modification, the device comprising: a first flow channel, an acoustic resonator capable of acting on liquid in the first flow channel, the acoustic resonator being in a shuttle shape, the acoustic resonator being used for generating acoustic streaming effect in the liquid in the first flow channel, so as to capture first particles in the first liquid, and make the captured first particles flow along the distribution of the shuttle-shaped contour of the acoustic resonator, so as to controllably mix the first particles distributed to the side edges of the shuttle-shaped contour with second liquid in the side edge area, and controllably mix so as to controllably combine second particles in the second liquid with the first particles; the first particles combined with the second particles flow to a fourth flow channel along the tail end of the shuttle-shaped contour of the acoustic resonator; and the first liquid after flowing through the first flow channel flows to two fifth flow channels.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical fields of micro-electro-mechanical, biomedicine, chemical industry, etc., and in particular, relates to a device and method for realizing particle modification. BACKGROUND

[0002] In the technical fields of medicine, biotechnology, chemical industry, etc., a scenario of modifying other target objects on a particle is usually encountered, for example, a particle is used as a carrier to modify other particles. However, how to improve the controllability of the modification process, improve the modification effect (such as high uniformity, etc.), and conveniently separate the modified particles to improve the overall modification process efficiency is a technical problem that has been concerned in research and development. The following takes the modification of nanoparticles (NPs) on red blood cells (RBC) as an example for illustration:

[0003] Red blood cells can be used as carriers for the delivery of drug-loaded nanoparticles. Among them, compared with free drugs, drug-loaded nanoparticles show significant advantages in targeting and sustained release, but they are easily cleared by the mononuclear phagocyte system in the spleen and liver, which leads to a decrease in the dose at the target site, thereby reducing their therapeutic effect. In order to solve this problem, loading (i.e., modifying) drug-loaded NPs onto the surface of red blood cells can make the drug circulate in the blood for a long time, thereby improving the drug delivery efficiency, effectively prolonging the circulation time of drug-loaded NPs in the body, and solving the common problem of premature clearance in drug-loaded NP delivery. This form of drug administration is also known as "red blood cell hitchhiking".

[0004] At present, the implementation method for loading NPs on the surface of red blood cells is mostly to adjust the properties of NPs (including size, surface charge, surface structure, etc.) to change their interaction with the red blood cell membrane, thereby producing different adhesion forces and the number of adhered NPs to be loaded onto the surface of red blood cells. The process usually involves an incubation process (for modifying NPs and loading onto red blood cells) and multiple centrifugation processes (for removing excess unbound NPs to obtain modified NPs of red blood cells), the overall scheme is complex and time-consuming, controllability is poor, and the level of automation is low, resulting in low production efficiency.

[0005] Therefore, how to improve the controllability of the modification of other particles by the particles as carriers (such as the modification of drug-loaded NPs by red blood cells), reduce the overall complexity, and improve the production efficiency is a technical problem to be solved. SUMMARY

[0006] In view of the above problems of the prior art, the present application provides a device and method for realizing particle modification to solve the technical problems mentioned in the background.

[0007] To achieve the above object, the first aspect of the present application provides a device for realizing particle modification, comprising: a first flow channel; a second flow channel and two third flow channels connected to the upstream of the first flow channel, wherein the two third flow channels are located on both sides of the second flow channel, the second flow channel is used for passing a first liquid containing first particles, and the two third flow channels are used for passing a second liquid containing second particles, and the second liquid passed by the two third flow channels forms a sheath flow; a fourth flow channel and two fifth flow channels connected to the downstream of the first flow channel, wherein the two fifth flow channels are located on both sides of the fourth flow channel; an acoustic resonator capable of acting on the liquid in the first flow channel, the acoustic resonator is in a shape of a shuttle, the tips of the front end and the rear end of the acoustic resonator are respectively directed to the directions of the second flow channel and the fourth flow channel; the acoustic resonator is used for generating acoustic streaming effect in the liquid in the first flow channel, so as to capture the first particles in the first liquid, make the captured first particles flow along the distribution of the shuttle-shaped contour of the acoustic resonator, and make the first particles distributed to the side edges of the shuttle-shaped contour controllably mixed with the second liquid in the side edge region, so as to controllably combine the second particles in the second liquid with the first particles; the first particles combined with the second particles flow to the fourth flow channel along the tip of the rear end of the shuttle-shaped contour of the acoustic resonator; and the first liquid after flowing through the first flow channel flows to the two fifth flow channels.

[0008] Optionally, the diameter of the inlet of the fourth flow channel is smaller than the diameter of the inlet of the fifth flow channel.

[0009] Optionally, the device further comprises: adjusting the pressure of the fourth flow channel to control the flow distribution between the fourth flow channel and the two fifth flow channels, so as to facilitate the first particles combined with the second particles to flow into the fourth flow channel and facilitate the first liquid after flowing through the first flow channel to flow to the two fifth flow channels.

[0010] Optionally, the device further comprises at least one of the following: a first inlet connected to the upstream of the second flow channel; a second inlet connected to the upstream of the two third flow channels; a third outlet connected to the downstream of the fourth flow channel; and a fourth outlet connected to the downstream of the two fifth flow channels.

[0011] Optionally, the height of the first flow channel and the power of the acoustic resonator are selected in relation to the first particles and the second particles.

[0012] Optionally, the particle size of the first particles is larger than the particle size of the second particles; under the current height of the first flow channel, the power of the acoustic resonator is controlled to capture the first particles, make the captured first particles flow along the distribution of the shuttle-shaped contour of the acoustic resonator, and make the first particles combined with the second particles flow to the downstream fourth flow channel at the tip of the rear end; and the power of the acoustic resonator is controlled to make the second particles not flow to the downstream at the tip of the rear end.

[0013] Optionally, the acoustic streaming effect generated by the acoustic resonator also induces the second particles to flow along the periphery of the acoustic resonator's shuttle profile, so as to controllably mix the first particles distributed to the side edges of the shuttle profile with the second liquid flowing along the periphery of the shuttle profile.

[0014] Optionally, the controllable mixing is achieved by at least one of the following: by controlling the power of the acoustic resonator; by controlling the flow rate of the first liquid, the flow rate of the second liquid, the flow rate ratio of the first liquid to the second liquid, or the total flow rate of the first liquid and the second liquid.

[0015] Optionally, the first particles comprise carriers, and the carriers comprise at least one of the following: cells, red blood cells, T cells, NK cells, stem cells, liposomes, microspheres; and the second particles comprise nano- or micro-particles, and the nano- or micro-particles comprise at least one of the following: drug-loaded nanoparticles, polymeric nanoparticles, inorganic nanoparticles, metallic nanoparticles, composite nanoparticles, magnetic nanoparticles, proteins, polypeptides, molecules, supramolecules.

[0016] The second aspect of the present application provides a method for implementing particle modification, using the device for implementing particle modification according to any one of the first aspect, the method comprising: introducing a first liquid containing first particles into a second flow channel; introducing a second liquid containing second particles into two third flow channels, the second liquid introduced by the two third flow channels forming a sheath flow; driving the acoustic resonator to work, generating an acoustic streaming effect in the liquid in the first flow channel, capturing the first particles in the first liquid based on the acoustic streaming effect, and making the captured first particles flow along the periphery of the acoustic resonator's shuttle profile, so as to controllably mix the first particles distributed to the side edges of the shuttle profile with the second liquid in the side edge region, so as to achieve the combination of the second particles in the second liquid with the first particles; the first particles combined with the second particles flow along the rear end tip of the acoustic resonator's shuttle profile to a fourth flow channel; and the first liquid after flowing through the first flow channel flows to two fifth flow channels.

[0017] Wherein, the scheme of the embodiment of the application can be applied to loading NPs to the surface of RBCs. For example, RBCs and 200 nm positively charged fluorescent particles are selected as the experimental model. The surface of RBCs is negatively charged, while the surface of NPs is positively charged. Due to electrostatic interaction, this promotes their combination. In this embodiment, the distribution and movement of RBCs and NPs under the acoustic streaming effect are studied, and the appropriate flow rate and input power are determined to control the NPs loading process. By adjusting the contact time between red blood cells and NPs, the loading amount of NPs can be adjusted to ensure uniform and controllable loading of NPs on the surface of red blood cells. On the other hand, in the embodiment of the application, the size and structure of the microchannel are designed to match the resonator, so that the modified NPs can be directly obtained after the RBC sample is modified, eliminating the repeated centrifugal washing steps required in the traditional in-phase method. The scheme of the embodiment has the characteristics of high uniformity of the modified NPs of RBCs, controllability and automation of the modification process, etc. BRIEF DESCRIPTION OF DRAWINGS

[0018] Figure 1 is a schematic diagram of the device structure for realizing particle modification provided by the first embodiment of the application;

[0019] Figure 2 is a diagram of the trapping effect of two kinds of particles under different channel heights and powers provided by the embodiment of the application, wherein (a) is a diagram of the trapping effect of 200 nm particles under different channel heights and powers, and (b) is a diagram of the trapping effect of 5 μm particles under different channel heights and powers;

[0020] Figure 3 is a schematic diagram of the characterization of particle manipulation provided by the embodiment of the application, wherein (a) is a schematic diagram of the microchannel structure, (i) is a local enlarged view of the position in the red dashed line frame, and (ii) is a top view of the assembled resonator and microchannel, (b) is a distribution diagram of two kinds of particles under different powers, and (c) is a result diagram of the NPs removal rate, wherein the vertical axis represents the concentration ratio between the fourth outlet and the second inlet, and the horizontal axis represents the input power of the resonator;

[0021] Figure 4 is a comparison diagram provided by the embodiment of the application, wherein (a), (b) and (c) are flow cytometry diagrams of the microfluidic and incubation methods under different particle concentrations (from left to right: 0.04 mg / mL, 0.1 mg / mL and 0.2 mg / mL), (d) is a loading efficiency diagram of nanoparticles (concentration: 0.04 mg / mL) under different input powers, (e) is a diagram of the percentage of positive cells, and (f) is a diagram of the statistical data of fluorescence intensity;

[0022] Figure 5is a biocompatibility characterization provided by an embodiment of the present application, wherein (a) shows CLSM images of NPs-loaded red blood cells shown in (i) bright field, (ii) red blood cell fluorescence field, (iii) NPs, and (iv) combination (captured at a power of 1360 mW and a concentration of NPs of 0.2 mg / mL), (b) is a confocal layer scanning image of a single red blood cell, (c) is a SEM image of natural red blood cells and NPs-loaded red blood cells, and (d) is a hemolysis result graph at different input powers;

[0023] Figure 6 is a schematic diagram of an apparatus structure for implementing particle modification provided by another embodiment of the present application.

[0024] It should be understood that in the above schematic diagram, the size and shape of each block diagram are only for reference, and should not constitute an exclusive interpretation of the embodiments of the present application. The relative position and inclusion relationship between the block diagrams presented by the schematic diagram only represent the structural association between the block diagrams, and are not limited to the physical connection mode of the embodiments of the present application. DETAILED DESCRIPTION

[0025] The technical solutions provided by the present application will be further described below in combination with the drawings and embodiments. It should be understood that the system structure and business scenarios provided in the embodiments of the present application are mainly to illustrate possible implementation modes of the technical solutions of the present application, and should not be interpreted as the only limitation of the technical solutions of the present application. Those skilled in the art can know that the technical solutions provided by the present application are also applicable to similar technical problems as the system structure evolves and new business scenarios appear.

[0026] It should be understood that the scheme for implementing particle modification provided by the embodiments of the present application includes an apparatus and a method for implementing particle modification, and an application thereof. Since the principles of solving problems of these technical solutions are the same or similar, in the introduction of the following specific embodiments, some repeated parts may not be described again, but should be regarded as mutual reference between these specific embodiments, which can be combined with each other.

[0027] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present application belongs. If there is any inconsistency, the meaning explained in the specification or the meaning derived from the content described in the specification shall prevail. In addition, the terms used herein are only for the purpose of describing the embodiments of the present application and are not intended to limit the present application. In order to accurately describe the technical content in the present application, and in order to accurately understand the present application, before the specific embodiments are described, the terms used in the specification are first explained as follows:

[0028] 1) Sheath flow focusing, a way of flow cytometry fluid system by using hydrodynamic focusing of cells in liquid, the principle is to use the faster moving sheath liquid to force the sample into a smaller core flow (also known as hydrodynamic core, namely the laminar flow near the axis), each particle in the core flow travels along the same axis at approximately the same speed. After the sample enters the core flow, it can be detected by a laser detection point.

[0029] Two-dimensional sheath flow focusing refers to the sheath flow focusing function based on micro-electro-mechanical system (MEMS), such as micro-channel based. Since the sheath flow and the sample flow are in the plane, it is called two-dimensional sheath flow focusing.

[0030] It should be noted that the structure that can form a laminar flow in the micro-channel in the present application is collectively referred to as a two-dimensional sheath flow structure.

[0031] 2) Acoustic resonator: refers to a resonator that can emit sound waves, wherein the vibration frequency of the acoustic resonator in the present application is about 0.5-30GHz, preferably about 1-10GHz, and more preferably about 1-5GHz. Among them, the acoustic resonator can be a bulk acoustic wave resonator (BAW, Bulk Acoustic Wave), a surface acoustic wave resonator (SAW, Surface Acoustic Wave), etc. The bulk acoustic wave resonator has a higher vibration frequency than the surface acoustic wave resonator.

[0032] Bulk acoustic wave resonator: under the excitation of an input signal, it generates ultrahigh frequency vibration, emits bulk acoustic waves, and the bulk acoustic waves propagate in the fluid to induce directional motion of the fluid, forming fluid vortex. One phenomenon is that several small fluid vortices can be generated at the edge of the chip profile of the bulk acoustic wave resonator in the fluid, and these fluid vortices can capture particles in the liquid. Combined with the position distribution of these fluid vortices, the phenomenon of the distribution of particles in the liquid at the edge of the chip profile of the bulk acoustic wave resonator is presented. Since the particles in the fluid flow through these fluid vortices distributed at the edge of the chip profile, the flow route of the particles formed by these fluid vortices is referred to as an acoustic fluid tunnel. The particles flowing through the acoustic fluid tunnel under the joint action of the fluid and the vortex can have a spiral-shaped travel route.

[0033] The first embodiment of the present application provides a device for realizing particle modification, which can be seen from the example shown in a of Figure 1 or Figure 6 The device can include:

[0034] A first flow channel;

[0035] a second flow channel and two third flow channels connected to the upstream of the first flow channel, wherein the two third flow channels are located on both sides of the second flow channel, the second flow channel is used to pass the first liquid containing the first particles, and the two third flow channels are used to pass the second liquid containing the second particles, and the second liquid passed by the two third flow channels forms a sheath flow;

[0036] a fourth flow channel and two fifth flow channels connected to the downstream of the first flow channel, wherein the two fifth flow channels are located on both sides of the fourth flow channel;

[0037] an acoustic resonator capable of acting on the liquid in the first flow channel, the acoustic resonator is in a shape of a shuttle, and the tips of the front end and the rear end of the acoustic resonator are respectively directed towards the second flow channel and the fourth flow channel;

[0038] the acoustic resonator is used to generate an acoustic streaming effect in the liquid in the first flow channel, so as to capture the first particles in the first liquid, and make the captured first particles flow along the distribution of the shuttle-shaped contour of the acoustic resonator, so as to realize the controllable mixing of the second particles in the second liquid with the first particles distributed to the side edges of the shuttle-shaped contour, and the controllable mixing is used to realize the controllable combination of the second particles in the second liquid with the first particles;

[0039] the first particles combined with the second particles flow to the fourth flow channel along the tip of the rear end of the shuttle-shaped contour of the acoustic resonator, and the first liquid after flowing through the first flow channel flows to the two fifth flow channels.

[0040] In some embodiments, the diameter of the inlet of the fourth flow channel is smaller than the diameter of the inlet of the fifth flow channel. Since the first particles combined with the second particles are released downstream along the tip of the rear end of the acoustic resonator, the diameter of the inlet of the fourth flow channel opposite to the tip of the rear end of the acoustic resonator can be smaller, and the diameter of the inlet of the fifth flow channel can be larger, so as to facilitate the liquid containing the unreacted second particles to carry the second particles to the fifth flow channel.

[0041] In some embodiments, the pressure of the fourth flow channel can be adjusted to control the flow distribution between the fourth flow channel and the two fifth flow channels, so as to facilitate the first particles combined with the second particles to flow into the fourth flow channel, and facilitate the first liquid after flowing through the first flow channel to flow to the two fifth flow channels. In some embodiments, the third outlet of the fourth flow channel is connected with a flexible pipe, and the flexible pipe is provided with a flow adjusting device (such as an adjustable clamping device) to adjust the pressure of the fourth flow channel.

[0042] In some embodiments, as shown in Figure 1 the second flow channel is connected with a first inlet upstream thereof, the two third flow channels are connected with a second inlet upstream thereof, the fourth flow channel is connected with a third outlet downstream thereof, and the two fifth flow channels are connected with a fourth outlet downstream thereof.

[0043] In some embodiments, the height of the first flow channel, the power of the acoustic resonator, and the first and second particles are selected in relation to each other. In some embodiments, the acoustic resonator has different trapping capabilities for the first and second particles at a certain height of the first flow channel and power of the acoustic resonator, e.g., the acoustic resonator has a stronger trapping capability for the first particles, or the first particles bound to the second particles, than for the second particles.

[0044] In some embodiments, the first particles have a larger particle size than the second particles; the power of the acoustic resonator is controlled so that it can trap the first particles at the current height of the first flow channel, the trapped first particles flow along the acoustic resonator's shuttle-shaped profile and bind to the second particles, and the first particles bound to the second particles flow downstream to the fourth flow channel at the tip of the back end; and the power of the acoustic resonator is controlled so that the second particles are not concentrated at the tip of the back end to flow downstream. That is, the acoustic resonator at this power has a stronger trapping capability for the first particles, or the first particles bound to the second particles, than for the second particles.

[0045] In some embodiments, the acoustic resonator's acoustic streaming effect also induces the second particles to flow along the periphery of the acoustic resonator's shuttle-shaped profile, so that the first particles distributed to the sides of the shuttle-shaped profile controllably mix with the second liquid flowing along the periphery of the shuttle-shaped profile in the side regions. The acoustic resonator at this power has a stronger trapping capability for the first particles, or the first particles bound to the second particles, than for the second particles, so the second particles are distributed to the periphery of the profile more than the first particles. Since the acoustic resonator also induces the second particles, it makes the second particles also flow along the periphery of the acoustic resonator's shuttle-shaped profile, increasing the probability of contact and collision with the first particles, and enhancing the binding effect.

[0046] In some embodiments, the controllable mixing is achieved by at least one of the following: by controlling the power of the acoustic resonator; by controlling the flow rate of the first liquid, the flow rate of the second liquid, the flow rate ratio of the first liquid to the second liquid, or the total flow rate of the first liquid and the second liquid.

[0047] In some embodiments, the first particles include carriers, and the carriers include at least one of the following: cells, red blood cells, T cells, NK cells, stem cells, liposomes, microspheres; and the second particles include nanoparticles, and the nanoparticles include at least one of the following: drug-loaded nanoparticles, polymer nanoparticles, inorganic nanoparticles, metal nanoparticles, composite nanoparticles, magnetic nanoparticles, proteins, polypeptides, molecules, supramolecules.

[0048] In some embodiments, the nanoparticles described above can also be microparticles. In some embodiments, the carrier-modified particles described above can be, for example, drug-loaded nanoparticles modified on cells (such as red blood cells, T cells, etc.), small molecules modified on the surface of cells, microspheres modified with proteins or polypeptides on the surface, and the like.

[0049] In some embodiments, the acoustic resonator comprises a single bulk acoustic wave resonator, a single surface acoustic wave resonator, or a combination of two or more acoustic resonators, such as a combination of two bulk acoustic wave resonators, or a combination of a bulk acoustic wave resonator and a surface acoustic wave resonator. The combination forms described herein include parallel combination or series combination.

[0050] For example, the parallel combination includes arranging the acoustic resonators side by side in the first flow channel, in which the width of the first flow channel can accommodate the side-by-side acoustic resonators, and such a structure can achieve a higher flux. Correspondingly, a plurality of fourth flow channels corresponding to the back ends of the plurality of acoustic resonators can be provided, and these fourth flow channels can be merged downstream into an outlet. Alternatively, another acoustic resonator for capturing particles can be arranged downstream of the first flow channel, to capture the first particles combined with the second particles flowing upstream, and then release the captured particles to the corresponding fourth flow channel through the other acoustic resonator.

[0051] For example, the series combination includes arranging the acoustic resonators in sequence upstream and downstream in the first flow channel, in which the length of the first flow channel can accommodate the series acoustic resonators, and such a structure can achieve a controllable reaction through the plurality of acoustic resonators in sequence, can prolong the controllable mixing reaction time, and can achieve the mixing reaction for a desired time. According to actual needs, there can be multiple parallel combinations and multiple series combinations, or a combination of parallel and series combinations.

[0052] For example, a plurality of acoustic resonators arranged in sequence along the extension direction of the first flow channel can further prolong the reaction time of the first liquid and the second liquid.

[0053] In some embodiments, as shown in b of Figure 6 , the third flow channel connected to the upstream of the first flow channel and the fifth flow channel connected to the downstream of the first flow channel can also be one, in which the third flow channel forms a single-sided sheath flow. In this scheme, the utilization rate of the acoustic resonator is lower than that of the scheme of a of Figure 1 or Figure 6 .

[0054] In some embodiments, the acoustic resonator is disposed on the channel wall of the first flow path, and the position thereof covers the first liquid and the second liquid, and the mixing of the two liquids is completed in the area covered by the acoustic resonator. For example, the front end of the profile of the acoustic resonator is located on the channel wall of the fluid path, and the position thereof covers the first liquid, and at least one side edge is located on the channel wall of the fluid path, and the position thereof covers the second liquid, so as to achieve the controllable mixing of the first liquid induced into the second liquid of the sheath flow layer. As shown in Figure 6 Fig. 2a shows an embodiment in which a part of the two side edges of the acoustic resonator extends into the area of the laminar flow of the sheath flow (i.e. the second liquid), so as to induce the first liquid in the laminar flow to flow along the two side edges and to react with the second liquid of the two sheath flow layers in a controllable manner. As shown in Figure 6 Fig. 2b shows another embodiment in which two-dimensional sheath flow and the first liquid form two laminar flows, and the first liquid is limited to one side of the laminar flow of the sheath flow (i.e. the laminar flow formed by the second liquid). The front end of the profile of the acoustic resonator is located in the area of the laminar flow of the first liquid, and a part of one side edge of the acoustic resonator extends into the area of the laminar flow of the sheath flow (i.e. the second liquid), so as to induce the first liquid in the laminar flow to flow along the side edge and to react with the second liquid of the sheath flow layer in a controllable manner.

[0055] In some embodiments, the vibration frequency of the acoustic resonator is about 0.5-30 GHz, preferably about 1-10 GHz, and more preferably about 1-5 GHz. When the vibration frequency is above 1 GHz, the effect of inducing the first liquid in the laminar flow to flow along the side edges of the profile of the acoustic resonator is very obvious, and the thermal effect is low. When the vibration frequency is below 5 GHz, it is relatively easy to control, so the above-mentioned more preferable range is about 1-5 GHz. On the other hand, the bulk acoustic resonator has a higher vibration frequency than the surface acoustic wave resonator, so the bulk acoustic resonator can be preferably used when a higher vibration frequency is required.

[0056] Preferably, the acoustic resonator is in the shape of a shuttle, for example, a shuttle with smooth side edges includes an angular front end, two side edges extending along the two sides of the angle, and a rear end formed by the closure of the two side edges at the downstream side to form a pointed end. In other embodiments, the acoustic resonator with a pointed or acute-angled front end and rear end, such as a rhombus, a polygon similar to a shuttle, a spindle structure, etc., can also be applied to the embodiments of the present application.

[0057] The second embodiment of the present application provides a method for realizing particle modification, which uses any of the above-mentioned devices for realizing particle modification, and the method comprises:

[0058] passing the first liquid containing the first particles into the second flow path;

[0059] passing the second liquid containing the second particles into two third flow paths, and the second liquid passed into the two third flow paths forms a sheath flow;

[0060] driving the acoustic resonator to operate, generating acoustic streaming effect in the liquid in the first flow channel, capturing first particles in the first liquid based on the acoustic streaming effect, making the captured first particles flow along the acoustic resonator shuttle-shaped profile distribution, controllably mixing the first particles distributed to the side edges of the shuttle-shaped profile with the second liquid in the side edge area to achieve the combination of the second particles in the second liquid with the first particles;

[0061] The first particles combined with the second particles flow to the fourth flow channel along the rear end tip of the acoustic resonator shuttle-shaped profile; and the first liquid after flowing through the first flow channel flows to two fifth flow channels.

[0062] In some embodiments, the controllable mixing of the first particles and the second particles is achieved by controlling the power of the acoustic resonator, the flow rate of the first liquid, the flow rate of the second liquid, the flow rate ratio of the first liquid to the second liquid, or the total flow rate of the first liquid and the second liquid, thereby achieving the controllable combination (modification) of the second particles to the first particles.

[0063] In order to better understand the present application, the experimental process of the third embodiment provided in the present application is described in detail below. In this embodiment, red blood cells (RBCs) are used as the first particles, and drug-loaded NPs are used as the second particles, and the NPs are modified to the RBCs through the scheme of the present application.

[0064] In this embodiment, the proposed acoustic streaming platform (i.e. Figure 1 The device shown not only achieves the loading of NPs to the surface of RBCs, but also integrates the structure for removing unbound NPs into one device. The working mechanism of the acoustic resonator is that high-frequency vibration reduces the attenuation distance of sound waves in the liquid, resulting in rapid attenuation inside the liquid, thereby increasing the body force to drive fluid flow, forming acoustic streaming effect. At the same time, due to the constriction effect of the microchannel structure, the liquid in the microchannel presents a vortex flow pattern. At the same time, the pressure applied at the inlet generates velocity along the positive x-axis, eventually resulting in spiral flow of the liquid in the microchannel. Studies have shown that cells or particles flowing through the microchannel are affected by both acoustic radiation force and resistance caused by the acoustic field. The strength of the acoustic radiation force acting on the particles is closely related to their size, and larger particles bear greater force. This size-dependent force leads to different distribution patterns of particles of different sizes in the microchannel. Therefore, the unique edge orientation of the resonator can control the distribution and direction of fluid and objects.

[0065] The PDMS microchannel is designed with two inlets and two outlets Figure 1). The second inlet is split into two branches for the sheath flow of NPs, while the first inlet is used for injection of the pre-treated RBC suspension. The third outlet serves as the collection port for the modified RBC sample, and the fourth outlet combines the two outlet branches into one for the outflow of excess free NPs. With this configuration, we achieved the direct collection of the red blood cell sample from the third outlet while transferring the excess unbound NPs to the fourth outlet. This allowed us to achieve the loading of NPs onto the surface of RBCs without employing the centrifugation mentioned in the background art.

[0066] In the low Reynolds number microfluidic system, the RBCs and NPs suspension forms a three-phase laminar flow device working region, with red blood cells occupying the middle position phase and NP sheath flow remaining on both sides. When entering the acoustic resonator, the difference in size between the first particle and the second particle and the location of the first flow channel, they exhibit different behaviors in the microchannel. Larger red blood cells are subjected to greater acoustic radiation force in the resonance region, causing them to exhibit spiral flow along the edge of the acoustic resonator and eventually be released at the end of the acoustic resonator. On the other hand, smaller NPs are subjected to very small acoustic radiation forces, causing them to move with the fluid. In the process, NPs adhere to the surface of RBCs through electrostatic interaction. In addition, the number of rotations of red blood cells along the edge of the device determines the length of the movement path, which can be adjusted by changing the input power of the acoustic resonator. Thus, the contact time between red blood cells and NPs is adjusted, and the loading amount of NPs on red blood cells is controlled.

[0067] The experimental process of the embodiment will be described in detail below:

[0068] I. Introduction of the experimental process

[0069] 1.1. Determine the parameters of the microfluidic chip

[0070] First, determine the "fixed" geometric parameters (first flow channel height) using simulation, and then determine the variable parameters such as power and flow rate using experiments.

[0071] On this platform, we predict that acoustic streaming will exert stronger manipulation on larger size particles and move them along the edge of the device. At the same time, the capture of smaller particles is minimized, ensuring that a large number of larger particles can flow out from the third outlet while minimizing the contamination of small particles. Therefore, the RBC sample loaded with nanoparticles can be directly collected from the third outlet without the need for further purification steps such as centrifugation. Studies have shown that for the acoustic streaming platform, the input power and the height of the microchannel are the main factors that determine whether particles will be captured at the edge of the device. Therefore, in order to determine the optimal height of the microchannel (i.e. the first flow channel) of our microfluidic chip, we studied the movement and distribution of particles in microchannels with heights of 20, 40, 80 and 100 μm under different input power conditions. Figure 2Figure a shows the results for 200 nm particles in microchannels at different heights, with power ranging from 460 mW to 1660 mW. 200 nm particle suspensions were introduced into both the first and second inlets, with a flow rate of 5 μL / min at the second inlet and 0.5 μL / min at the first inlet. It is shown that decreasing the microchannel height and increasing the input power can enhance particle trapping. For example, the trapping effect of 200 nm particles in a 20 μm microchannel is very significant. However, when the microchannel height is 40 μm and the power is below 760 mW, or when the height is 80 μm or 100 μm and the power is below 1660 mW, there is no significant trapping of 200 nm particles. Therefore, we further investigated microchannels with heights of 40, 80, and 100 μm, for a total of 5 μm particles. Figure 2 As shown in Figure b, ultrapure water (UP water) was injected at a flow rate of 5 μL / min through the second inlet, while a 5 μm particle suspension was injected at a flow rate of 0.5 μL / min through the first inlet. However, for the 40 μm high microchannel, although there was no significant capture of 200 nm particles at low power (460 mW), the capture of 5 μm particles was also unsatisfactory. Similarly, for the 100 μm high microchannel, there was no significant capture of 200 nm particles when the power was below 1660 mW, but there was no effect on the manipulation of 5 μm particles. After comprehensive consideration, we finally determined to use a microchannel height of 80 μm and power of 760, 1060, and 1360 mW for further research, because these conditions can effectively capture 5 μm particles, while there is almost no significant capture of 200 nm particles. It can be seen that the conditions for particle size capture are related to the height of the microchannel (i.e., the first channel) and the input power of the acoustic resonator. Therefore, when it is necessary to capture particles of different sizes, the microchannel height and input power conditions for capturing particles of each size can be obtained through experiments, and then a suitable microchannel height and input power can be selected.

[0072] 1.2. Characterization of Sample Collection

[0073] To investigate the platform's performance in recovering large particles while removing small particles, polystyrene spheres with diameters of 5 μm and 200 nm were first used as model particles in the experiment. Figure 3 As shown in Figure a, suspensions of 200 nm and 5 μm polystyrene microspheres were injected from the second inlet and the first inlet, respectively. The third outlet was used to collect 5 μm particles, and the fourth outlet was used to collect 200 nm particles. During the experiment, the height of the collection tube was adjusted to regulate the pressure at the outlet, thereby controlling the flow distribution between the two outlets to ensure that as many 5 μm particles as possible flowed out from the third outlet and 200 nm particles flowed out from the fourth outlet. A video snapshot shows the particle flow distribution under different input powers in the resonator's operating region. Figure 3b) where orange is 5 pm particles and green is 200 nm particles. The results show that when the device is off, the 200 nm particle suspension on both sides is sandwiched in the middle 5 pm particle suspension, forming a three-phase laminar flow. In this case, almost all 5 pm particles can flow out from the middle outlet, while 200 nm particles flow out from the side branches. Once the device is activated, the trajectories of the particles change significantly due to the acoustic streaming effect. After entering the working region, 5 pm particles spiral along the edge of the resonance region first, then converge to the middle branch, and finally flow out through the third outlet. At the same time, 200 nm particles also flow spirally with the fluid, flow into the side branch, and flow out through the fourth outlet. The spiral motion path of 5 pm particles enhances their interaction with 200 nm particles.

[0074] In addition, we also calculated the concentration ratio between the fourth outlet and the second inlet. This ratio is defined as the 200 nm particle removal rate of the platform (details of the calculation in the experimental section). The results show that the removal rate is as high as 90% or more under all power conditions Figure 3 c) of FIG. 1. In summary, two types of particles processed by acoustic streaming flow out from different outlets without the need for post-processing like centrifugation.

[0075] 1.3. Loading NPs onto RBCs

[0076] To demonstrate this concept, we studied the loading of NPs on the surface of RBCs based on the flow parameters obtained from the above particle experiments. We injected a suspension of 200 nm positively charged NPs at a rate of 5 pL / min into the second inlet and a suspension of RBCs at a rate of 0.5 pL / min into the first inlet, respectively. Figure 4 a-c of FIG. 3 show the flow cytometry fluorescence intensity distribution obtained using the microfluidic method (blue histogram) and the batch incubation method (red histogram) for NPs concentrations of 0.04, 0.1, and 0.2 mg / mL, respectively. The results show that the fluorescence distribution of NPs-modified red blood cells obtained by the microfluidic method is narrower but the fluorescence intensity is relatively lower. This indicates that the microfluidic-based loading method effectively enhances the uniformity of the product, while the loading capacity is slightly reduced.

[0077] To study the effect of input power on the loading of NPs onto RBCs, we analyzed the loading efficiency of NPs at different power conditions (0 mW, 760 mW, 1060 mW, and 1360 mW) for a NPs concentration of 0.04 mg / mL. In these experiments, we only adjusted the input power of the resonator while keeping other experimental conditions unchanged. As shown in Figure 4As shown in the diagram, flow cytometry results indicate that the loading efficiency of RBCs increases with increasing power. Furthermore, the fluorescence intensity of RBCs modified with NPs increases with increasing power. Figure 4 (f) This indicates that the loading amount of NPs can be easily adjusted by controlling the input power of the resonator. We analyze that this is because the increase in input power leads to an increase in acoustic flow velocity. Furthermore, since the RBC and NP exhibit helical flow in the working region of the device, this lengthens the contact path between them as they flow through the resonant region at a fixed length, thus providing more opportunities for the particles to assemble onto the RBC surface. Figure 4g shows snapshots of the movement of 5-micron particles at power levels of 760, 1060, and 1360 mW. Statistical analysis of the number of rotations the particle takes to move from point A to point B reveals that the number of rotations within a certain distance increases with increasing power. This demonstrates that the length of the path the particle travels in the microchannel within the platform can be adjusted by power. This further demonstrates the controllability of the loading amount of red blood cell nanoparticles in this platform.

[0078] To verify the platform's biocompatibility, we visualized erythrocytes loaded with nanoparticles using confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). Here, the erythrocyte membrane was stained to achieve three-dimensional visualization through confocal imaging. Figure 5 As shown in the images, both CLSM and SEM images confirmed that the NPs were effectively loaded onto the RBCs. Simultaneously, the RBCs largely retained their natural structure, exhibiting a concave disc shape with a diameter of approximately 6-8 μm. This indicates that this method causes minimal damage to the RBCs. Furthermore, the hemolysis rate of RBC samples treated under different power conditions was further measured using a microplate reader. After centrifuging the obtained RBC suspensions loaded with NPs, the hemoglobin uptake intensity in the supernatant was measured to calculate the hemolysis rate (details of the calculation are in the Materials and Methods section). The results confirmed that the hemolysis rate of the samples obtained in this experiment was all below 5%, indicating that this method did not cause significant damage to the RBCs.

[0079] In summary, we present a microfluidic-based strategy for loading NPs onto erythrocytes in this work. While there is potential for further improvements in loading efficiency, this microfluidic chip integration approach demonstrates excellent controllability, integration, and automation in RBC carrier preparation, effectively reducing contamination issues during processing.

[0080] II. Conclusion

[0081] In this study, we introduce a platform for surface loading of NPs onto RBCs by integrating high-frequency piezoelectric acoustic resonators into a microfluidic structure with multiple inlet and outlet branches. The resonators are used to modulate the flow distribution and interaction between RBCs and NPs in the microchannel. Compared to the traditional bulk incubation method, the microfluidic system enhances the control of uniformity of micro / nano-objects. In addition, by adjusting the input power of the resonators, especially under low particle concentration conditions, the loading of NPs on RBCs can be controlled. This allows the control of the contact opportunity and duration between RBCs and NPs. In addition, the unique edge direction of the resonator guides the RBCs to focus in the middle of the microchannel. This, combined with the matching design of the microfluidic size and structure, helps to collect the modified RBCs loaded with NPs directly from the designated sample outlet, while the excess free NPs are guided to the waste outlet, eliminating the need for a tedious post-processing step to obtain the final sample. This greatly improves the automation level of the RBC loading process. As a versatile platform technology, the proposed platform has the potential for various applications, including particle or cell modification and washing studies.

[0082] III. Introduction of the experimental section

[0083] 3.1. Reagents and materials

[0084] Fluorescein isothiocyanate (FITC)-labeled 200 nm polystyrene NPs with positively charged surface groups were purchased from Qilu Biotech Co., Ltd. (Xi'an, China). 5 pm red uniform polystyrene microspheres were purchased from Shanghai Maikelin Biochemical Co., Ltd. (Shanghai, China). 200 nm non-functionalized polystyrene microspheres were purchased from Shanghai Huizhi Biotechnology Co., Ltd. (Shanghai, China). Hank's buffer was purchased from Solabio Technology Co., Ltd. (Beijing, China). Red fluorescent cell membrane dye in the neurite growth staining kit was purchased from Thermo Fisher Scientific (USA). Glutaraldehyde fixative (2.5%) was purchased from Shanghai Yuanye Biotechnology Co., Ltd. (Shanghai, China). Analytical grade ethanol was purchased from Tianjin Yuanda Company (Tianjin, China). Ultra-pure water (UP water, Millipore filtration system, resistivity higher than 18.2 MΩ). Polydimethylsiloxane (PDMS, Sylgard 184) was purchased from Dow Chemical Company. Negative photoresist (SU-8) was purchased from Suzhou Scientific Research Material Micro-technology Co., Ltd. (Suzhou, China). All reagents were not further purified.

[0085] 3.2. Fabrication of the platform

[0086] The core components of the platform include a high-frequency acoustic resonator and a PDMS microfluidic structure with two inlets and two outlets. The resonator is fabricated using microelectromechanical systems (MEMS) technology. Briefly, alternating deposition of aluminum nitride (ALN) and silicon dioxide (Si02) on a silicon wafer forms a reflective layer. Molybdenum (Mo) is deposited as a bottom electrode, followed by ALN as a piezoelectric layer, and then another layer of Mo as a top electrode. Each layer is patterned accordingly through an etching process. This results in the formation of the “sandwich” structure of the resonator with the desired resonator area.

[0087] The microfluidic structure is fabricated using standard photolithography techniques. Briefly, the pattern of a photomask is first designed using Layout software and fabricated through mask photolithography. SU-82025 or SU-82050 negative photoresist is spin-coated onto a clean 4-inch silicon wafer with the desired thickness. Subsequently, pre-baking is performed on a hot plate, followed by transferring the patterned structure from the photomask to the photoresist using a UV photolithography machine. After photolithography, post-baking, and development, re-heating is performed to further strengthen the microstructure, resulting in a wafer with SU-8 photoresist structure. To replicate the microchannel structure, PDMS-based casting and curing agents are thoroughly mixed in a mass ratio of 10:1 and vacuumed to remove air bubbles. To ensure smoothness, the PDMS mixture is poured into the silicon wafer mold produced above and pre-cured at 90°C on a flat hot plate for 20 minutes. Then, additional curing is performed in a constant-temperature oven for 90 minutes to achieve full curing. Microchannel structures of different heights are obtained. It consists of two inlets and two outlets, with the main channel having a width of 500 pm, including three inlet branches and three outlet branches. The width of the inlet branches is 300 pm each. The width of the middle outlet branch is 60 pm, and the width of the outlet branches on both sides is 600 pm. Finally, after creating the liquid inlet and outlet holes, the PDMS microfluidic channel is assembled with the resonator using a 3D-printed jig.

[0088] 3.3. Particle experiments for evaluating the platform

[0089] The optimal microchannel height was investigated using surface-uncharged particles and the non-centrifugal ability of the platform was verified. 200 nm green fluorescent particles and 5 pm red fluorescent particles were injected from the second inlet and the first inlet, respectively, through polytetrafluoroethylene tubes. A fluorescence microscope was used to observe the flow and distribution of particles in real time. The sample concentration collected from the fourth outlet was measured by NTA (NS-300, Malvern, UK).

[0090] 3.4. Loading of NPs to RBCs

[0091] Whole blood was collected from healthy mice and centrifuged at 300 g for 3 times at 4 °C to wash the serum and resuspended in Hank’s buffer. By dilution with Hank’s buffer, different concentrations of positively surface charged NPs were obtained. For microfluidic method, positively surface charged NPs were injected from the second inlet into the microchannel at a flow rate of 5 pL / min, while red blood cells were injected from the first inlet into the microchannel at a flow rate of 0.5 pL / min. After acoustic streaming loading, NPs-loaded RBCs were collected from the third outlet using a centrifuge tube containing Hank’s buffer, while the excess NPs were flowed out from the fourth outlet. For incubation method, NPs and RBCs suspension were mixed and incubated at a volume ratio of 10:1, then Hank’s buffer was added and centrifuged for 3 times to wash the excess NPs. The obtained samples were stored at 4 °C for flow cytometry analysis.

[0092] 3.5. Flow cytometry

[0093] Flow cytometry (CytoFLEX LX, Beckman Coulter, USA) was used to analyze the fluorescence intensity and loading of NPs, using 488 nm laser for excitation and 525 nm emission filter for FITC detection. Before flow cytometry assay, the red blood cell sample was filtered through a 300-mesh filter to prevent instrument clogging. Data were processed using FlowJo 10.8.1.

[0094] 3.6. SEM characterization

[0095] After centrifugation of the native red blood cell and NPs-loaded red blood cell samples, they were fixed using 2.5% glutaraldehyde for 30 min. Subsequently, gradient dehydration was performed using 30%, 50%, 70%, 95%, and 100% ethanol for 10 min each. Finally, the red blood cells were resuspended in 95% ethanol. The prepared samples were spotted onto clean glass slides and left to dry naturally overnight. After gold sputtering, the prepared samples were imaged for SEM (Apreo S, Thermo Scientific, USA).

[0096] 3.7. Characterization of red blood cell hemolysis

[0097] The hemolysis rate of RBCs after treatment with NPs surface loading platform was determined using a microplate reader (Varioskan LUX Multimode Microplate Reader, Thermofisher, USA). First, the absorbance wavelength of hemoglobin was determined. A 2.5 pL sample of the original concentration RBC suspension was diluted in 1 mL of UP water. Then, it was refrigerated at 4°C for 1 hour, with gentle mixing every 20 minutes to ensure complete release of hemoglobin. After centrifugation (3000 g, 5 minutes, 4°C), the supernatant containing hemoglobin was obtained. Then 200 pL of the supernatant sample was aspirated and diluted twice before being added to a standard 96-well plate. The ultraviolet absorbance intensity was measured in the range of 350 nm to 750 nm to determine the peak absorbance wavelength, which was found to be 415 nm.

[0098] To measure the hemolysis level of the experimental group (EG), we collected the supernatant after centrifugation (300 g, 5 minutes, 4°C) of the RBC sample suspension treated with the platform. Each sample under different power conditions was aliquoted 200 pL into a 96-well plate for measurement of the ultraviolet absorbance intensity at 415 nm. Likewise, to measure the hemolysis level of the positive control group (PCG) and the negative control group (NCG), we suspended an equal amount of red blood cells in UP water to completely release hemoglobin, which theoretically would not hemolyze, but release hemoglobin in Hank's buffer. Then, we measured the absorbance peak at a wavelength of 415 nm using the same method. The hemolysis rate (HR%) was calculated using the following formula:

[0099]

[0100] where Absorbance values are the values of absorbance, EG is the experimental group, NCG is the negative control group, and PCG is the positive control group.

[0101] In several embodiments provided in the present application, it should be understood that the disclosed apparatus and methods are not limited to the above embodiments, and can also be implemented in other manners. For example, the above-described apparatus embodiments are merely schematic, and the division of the units is merely a logical function division, and actual implementation can have another division manner, for example, a plurality of units or components can be combined or integrated into another system, or some features can be ignored or not executed. In addition, the coupling or direct coupling or communication connection between the units shown or discussed can be indirect coupling or communication connection through some interfaces, apparatuses or units, and can be electrical, mechanical or other forms.

[0102] The units described as separate components may or may not be physically separate, and the components displayed as units may or may not be physical units, i.e. may be located in one place, or may also be distributed on multiple network units. Part or all of the units can be selected according to actual needs to achieve the purpose of the embodiment scheme.

[0103] In addition, the words "first", "second", "third" or "module A", "module B", "module C" and the like in the specification and claims are only used to distinguish similar objects, and do not represent a specific order of the objects. It can be understood that the specific order or sequence can be interchanged as permitted, so that the application described herein can be implemented in an order other than that illustrated or described herein.

[0104] In the above description, the labels indicating steps, such as S10, S20, …, etc., do not necessarily mean that the steps are executed in this order, and the order of the steps before and after can be interchanged or executed simultaneously as permitted.

[0105] The term "comprising" used in the specification and claims should not be interpreted as limited to the listed elements; it does not exclude other elements or steps. Therefore, it should be interpreted as specifying the presence of the stated features, integers, steps or components as referred to, but does not preclude the presence or addition of one or more other features, integers, steps, components or groups thereof. Therefore, the expression "a device comprising means A and B" should not be limited to a device consisting only of components A and B.

[0106] The term "one embodiment" or "an embodiment" mentioned in the specification means that the specific features, structures or characteristics described in connection with the embodiment are included in at least one embodiment of the application. Therefore, the phrase "in one embodiment" or "in an embodiment" appearing throughout the specification does not necessarily refer to the same embodiment, but can refer to the same embodiment. In addition, in one or more embodiments, each specific feature, structure or characteristic can be combined in any appropriate manner as apparent to those skilled in the art from the disclosure.

[0107] Note that the above is only the preferred embodiment of the application and the technical principles applied. Those skilled in the art will understand that the application is not limited to the specific embodiments described herein, and those skilled in the art can make various obvious changes, readjustments and substitutions without departing from the scope of the application. Therefore, although the application has been described in more detail through the above embodiments, the application is not limited to the above embodiments, and can include more other equivalent embodiments without departing from the concept of the application, all of which fall within the scope of the application.

Claims

1. An apparatus for implementing particle modification, characterized by, The device comprises: a first flow channel; a second flow channel and two third flow channels connected to the upstream of the first flow channel, wherein the two third flow channels are located on both sides of the second flow channel, the second flow channel is used to pass a first liquid containing first particles, and the two third flow channels are used to pass a second liquid containing second particles, and the second liquid passed by the two third flow channels forms a sheath flow; a fourth flow channel and two fifth flow channels connected to the downstream of the first flow channel, wherein the two fifth flow channels are located on both sides of the fourth flow channel; an acoustic resonator capable of acting on the liquid in the first flow channel, the acoustic resonator is in the shape of a shuttle, and the tips of the front end and the rear end of the acoustic resonator are respectively directed towards the second flow channel and the fourth flow channel; the particle size of the first particles is larger than the particle size of the second particles, so that when the height of the first flow channel and the power of the acoustic resonator are certain selected values, the trapping ability of the acoustic resonator for the first particles or the first particles combined with the second particles is stronger than the trapping ability of the acoustic resonator for the second particles; the acoustic resonator is used to generate an acoustic streaming effect in the liquid in the first flow channel, and the power of the acoustic resonator is controlled at the current height of the first flow channel to trap the first particles in the first liquid, so that the trapped first particles flow along the shuttle-shaped contour of the acoustic resonator, and the controllable mixing of the first particles distributed to the side edges of the shuttle-shaped contour with the second liquid in the side edge region of the acoustic resonator is achieved, so that the controllable combination of the second particles in the second liquid with the first particles is achieved; the controllable mixing is achieved by controlling the power of the acoustic resonator, wherein as the power increases, the number of revolutions of the first particles rotating within a certain distance along the shuttle-shaped contour of the acoustic resonator also increases, thereby increasing the length of the path of movement of the first particles and increasing the contact time between the first particles and each of the second particles, and the control of the loading amount of the second particles on the first particles is achieved; the first particles combined with the second particles flow to the fourth flow channel along the tip of the rear end of the shuttle-shaped contour of the acoustic resonator; the first liquid after flowing through the first flow channel flows to the two fifth flow channels; and the power of the acoustic resonator is such that the unreacted second particles are not concentrated at the tip of the rear end and flow to the downstream two fifth flow channels.

2. The apparatus of claim 1, wherein, The diameter of the inlet of the fourth flow channel is smaller than the diameter of the inlet of the fifth flow channel.

3. The apparatus of claim 2, wherein, Further comprising: adjusting the pressure of the fourth flow channel to control the flow distribution between the fourth flow channel and the two fifth flow channels, so as to facilitate the flow of the first particles combined with the second particles into the fourth flow channel and the flow of the first liquid after flowing through the first flow channel to the two fifth flow channels.

4. The apparatus of any one of claims 1-3, wherein, Further comprising at least one of: a first inlet connected to the upstream of the second flow channel; a second inlet connected to the upstream of the two third flow channels; a third outlet connected to the downstream of the fourth flow channel; a fourth outlet connected to the downstream of the two fifth flow channels.

5. The device according to claim 1, wherein the height of the first flow channel and the power of the acoustic resonator are selected in relation to the first particles and the second particles.

6. The apparatus of claim 1, wherein, The acoustic resonator generates acoustic streaming effect, which also induces the second particles to flow along the periphery of the acoustic resonator shuttle profile, so that the first particles distributed to the side of the shuttle profile controllably mix with the second liquid in the side region of the shuttle profile.

7. The apparatus of claim 1, wherein, The controllable mixing is achieved by controlling the flow rate of the first liquid, the flow rate of the second liquid, the flow rate ratio of the first liquid to the second liquid, or the total flow rate of the first liquid and the second liquid.

8. The apparatus of claim 1, wherein The first particles comprise carriers, and the carriers comprise at least one of the following: cells, liposomes, microspheres. The second particles comprise nano- or micro-particles, and the nano- or micro-particles comprise at least one of the following: drug-loaded nanoparticles, polymeric nanoparticles, inorganic nanoparticles, metallic nanoparticles, composite nanoparticles, magnetic nanoparticles.

9. The apparatus of claim 8, wherein, The cells comprise at least one of the following: red blood cells, T cells, NK cells, stem cells.

10. The apparatus of claim 1, wherein The first particles comprise carriers, and the carriers comprise at least one of the following: cells, liposomes, microspheres. The second particles comprise at least one of the following: proteins, polypeptides.

11. The apparatus of claim 1, wherein The first particles comprise carriers, and the carriers comprise at least one of the following: cells, liposomes, microspheres. The second particles comprise molecules.

12. The apparatus of claim 1, wherein The first particles comprise carriers, and the carriers comprise at least one of the following: cells, liposomes, microspheres. The second particles comprise supramolecules.

13. A method of effecting particle modification, characterized by, The method comprises using the apparatus for modifying particles according to any one of claims 1-12, and the method comprises: feeding the first liquid containing the first particles into the second flow channel; feeding the second liquid containing the second particles into two third flow channels, and the second liquid fed into the two third flow channels forms a sheath flow; driving the acoustic resonator to work, generating acoustic streaming effect in the liquid in the first flow channel, capturing the first particles in the first liquid based on the acoustic streaming effect, and making the captured first particles flow along the shuttle profile of the acoustic resonator, so that the first particles distributed to the side of the shuttle profile controllably mix with the second liquid in the side region of the shuttle profile, to achieve the combination of the second particles in the second liquid with the first particles; the first particles combined with the second particles flow to the fourth flow channel along the rear tip of the shuttle profile of the acoustic resonator; and the first liquid after flowing through the first flow channel flows to the two fifth flow channels.

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